We found that short-term culture medium and homogenate of casein-induced rat peritoneal polymorphonuclear leukocytes (PMN) markedly induced collagenase and prostaglandin E2 (PGE2) production by normal rat synovial cells and these effects were abrogated by anti-(rat interleukin-1 alpha) (IL-1 alpha) polyclonal antibodies. However, collagenase activity and PGE2 induced by recombinant rat IL-1 alpha were less than those induced by rat PMN culture medium. It was also proved by radioimmunoassay that rat PMN culture medium contains a relatively small amount of IL-1 alpha. The introduction of IL-1 alpha-deleted PMN culture medium and recombinant rat IL-1 alpha together into the synovial cell culture system revealed that IL-1 alpha deleted PMN culture medium has a significant enhancing activity on IL-1 alpha-induced synovial cell collagenase and PGE2 production. This new factor, which was shown to be a negatively charged protein of about 80 kDa, may have important roles in connective tissue destruction and chronic inflammation in diseases such as rheumatoid arthritis.